N9664
Polynucleotide Phosphorylase from Escherichia coli
Synonym(s):
Polyribonucleotide: 0rthophosphate Nucleotidyltransferase
recombinant
expressed in E. coli
form
solution
enzyme activity
≥250 units/mg protein
mol wt
230 kDa
shipped in
dry ice
storage temp.
−70°C
General description
Polynucleotide phosphorylase (PNPase) is required for cell viability and mRNA turnover in Escherichia coli.
Application
Polynucleotide phosphorylase (PNP) has been used in a study to show that spontaneous mutations resulting from replication errors are reduced in a PNP-deficient strain. It has also been used in a study to show that the absence of PNPase makes E. coli cells sensitive to UV, which suggests PNP has a role in survival of UV damage.
Biochem/physiol Actions
Polynucleotide phosphorylase from Escherichia coli functions as a exonuclease as well as a poly(A) polymerase and can add C and U nucleotides to poly(A) tails.
Physical form
Supplied as a solution in 20 mM Hepes buffer pH 7.9, 0.1 mM EDTA, 2 mM DTT, 12.5 mM MgCl2, 200 mM KCl, 21.4% (w/v) Glycerol
Other Notes
One unit will polymerize 1.0 μmole of ADP releasing 1.0 μmole of inorganic phosphate in 15 minutes, at pH 9.1 at 37 °C.
Regulatory Information
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W P Donovan et al.
Proceedings of the National Academy of Sciences of the United States of America, 83(1), 120-124 (1986-01-01)
The isolation of a temperature-sensitive allele of RNase II (rnb) by in vitro mutagenesis has permitted the demonstration that RNase II and polynucleotide phosphorylase (PNPase) are required for cell viability and mRNA turnover in Escherichia coli. Double-mutant strains carrying the
A J Carpousis
Biochemical Society transactions, 30(2), 150-155 (2002-05-31)
mRNA instability is an intrinsic property that permits timely changes in gene expression by limiting the lifetime of a transcript. The RNase e of Escherichia coli is a single-strand-specific endo-nuclease involved in the processing of rRNA and the degradation of
Purification and properties of polynucleotide phosphorylase from Escherichia coli.
Y Kimhi et al.
The Journal of biological chemistry, 243(2), 231-240 (1968-01-25)
Involvement of pnp in survival of UV radiation in Escherichia coli K-12
Rath, D., et al.
Annual Review of Microbiology, 128, 1196-1205 (2012)
H Soreq et al.
The Journal of biological chemistry, 252(19), 6885-6888 (1977-10-10)
A simple procedure for purifying polynucleotide phosphorylase from Escherichia coli cells by means of affinity chromatography on an RNA-Sepharose column is described. The purified enzyme preparation has a specific activity 3500-fold that of the crude extract and is essentially homogeneous
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